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Corresponding author: Vlad Dincă ( vlad.e.dinca@gmail.com ) Academic editor: Sven Erlacher
© 2018 Vlad Dincă, Levente Székely.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Dincă V, Székely L (2018) First record of Scopula orientalis (Alphéraky, 1876) (Lepidoptera, Geometridae) in Romania, at the northern limit of the Balkans. Nota Lepidopterologica 41(2): 189-197. https://doi.org/10.3897/nl.41.24316
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The geometrid moth Scopula orientalis (Alphéraky, 1876) has an apparently disjunct distribution in Europe, with local populations in the Balkans (Macedonia, Bulgaria and Albania where recently discovered), as well as in Ukraine and southern European Russia. In this study, based on morphological and mitochondrial DNA (cytochrome c oxidase subunit 1 – COI) data, we report the presence of S. orientalis in south-eastern Romania (Dobrogea), at the northernmost limit of the Balkans. The flight time (September) of the recorded specimen, is the latest seasonal record for the Balkans, supporting the presence of at least a partial second generation. The Romanian specimen represented a unique COI haplotype which is differentiated by seven mutations from its genetically closest population in Sivas province, Turkey. The presence of S. orientalis in Romania reduces the distributional gap between the known Balkan and Ukrainian populations and highlights the potential for a more widespread distribution that needs documentation based on directed studies.
The ornata species-group of the genus Scopula Schrank, 1802 (family Geometridae) is represented in Europe by nine species (
In this study, based on genitalia morphology and DNA data, we report the presence of S. orientalis in the Romanian fauna.
We examined the genitalic morphology of 24 specimens (1 Bulgarian, 1 Spanish, 2 Italian and 20 Romanian) of the ornata species-group (Suppl. material
The male and female genitalia were processed as follows: maceration in 10% potassium hydroxide (ca. six minutes at 90 °C), cleaning in distilled water, examination under a stereomicroscope and storage in tubes with glycerine. Photos of the genitalia were taken in 70% ethanol by using a digital camera attached to a stereomicroscope. Specimens were identified based on comparison with illustrations and text from
From the 16 specimens sequenced during this study, total genomic DNA was extracted using Chelex 100 resin, 100–200 mesh, sodium form (Biorad), under the following protocol: one leg was removed and introduced into 100 μl of Chelex 10% and 5 μl of Proteinase K (20 mg/ml) were added. The samples were incubated overnight at 55 °C and were subsequently incubated at 100 °C for 15 min. Samples were then centrifuged for 10 s at 3000 rpm. A 658-bp fragment near the 5´ end of COI was amplified by polymerase chain reaction using the primers LepF1 (5´-ATTCAACCAATCATAAAGATATTGG-3´) and LepR1 (5´- TAAACTTCTGGATGTCCAAAAAATCA-3´) (
Sequences were edited and aligned using GENEIOUS PRO 6.1.8 created by Biomatters (http://www.geneious.com/). The 16 sequences obtained in this study have been submitted to GenBank (Suppl. material
The DNA analyses included 90 cytochrome c oxidase subunit 1 (COI) sequences. Seventy-one sequences represented all species of the European ornata species-group available in GenBank (six out of nine species present in Europe) and originated from
For DNA-based identifications, a neighbour-joining (NJ) tree was constructed in MEGA7 (
The genitalic examination of 21 specimens (1 Bulgarian, 20 Romanian) of the ornata species-group indicated that 18 represented S. decorata, two S. ornata and one S. orientalis (Suppl. material
Material. S. orientalis, 1 ♂, Enisala, Tulcea county (northern Dobrogea), Romania, 4.ix.2015, 85m (44°53.01'N, 28°50.10'E), leg. & coll. L. Székely. Prep. genit. 2318/Dincă, COI GenBank accesion number MG786139. Wing span 22 mm (Fig.
Scopula orientalis (Fig.
Male genitalia elements of S. orientalis, S. decorata and S. ornata. a-c. Male genitalia of S. orientalis from Romania (Enisala fortress, Tulcea county, 4.ix.2015), prep. genit. 2318/Dincă. a. Genital armature. b. Aedeagus. c. Sternum A8. d. Sternum A8 of male S. decorata, Romania (Enisala fortress, Tulcea county, 10.vii.2013), prep. genit. 2405/Dincă. e. Sternum A8 of male S. ornata, Romania (Enisala fortress, Tulcea county, 24.viii.2016), prep. genit. 2417/Dincă. Scale bar is 0.5 mm.
The NJ analysis showed that each species of the ornata species group formed a well-supported clade (Fig.
The clade formed by S. orientalis included the Romanian specimen that was also identified based on genitalic morphology. This specimen represented a unique haplotype diverging by seven mutations (ca 1%) from conspecific specimens from Sivas province (Turkey) and by 15 mutations (ca 2.3%) from the specimen from Gümüşhane (Fig.
Scopula orientalis was described as a variety of S. decorata (Acidalia decorata var. orientalis), based on material originating from south-western European Russia (
The presence of S. orientalis in the Romanian fauna is not unexpected, as it narrows the gap between the known populations in the Balkans and those in Ukraine. Since only one record is available, we cannot completely rule out the possibility that our specimen represents a stray individual originating from other areas. However, this hypothesis is unlikely because S. orientalis is a weak flyer and we have not found any notes describing migratory behaviour. Furthermore, the habitat from Enisala fortress (Fig.
In the Balkans, the few confirmed records of this species originate from Macedonia, south-western Bulgaria and south-eastern Albania (
The data above show that S. orientalis is a species with very poorly known distribution in Europe, especially in the Balkans. The lack of data is probably due both to the local character of the species, but also to the fact that it has been overlooked due to its external similarity to other Scopula species, particularly S. ornata and S. decorata.
The larval food plant of S. orientalis is unknown and detailed information on habitat preferences is very limited (
Collecting site of Scopula orientalis in Romania. a. Rocky steppe slopes at Enisala fortress, Tulcea county (photo taken on 30.ix.2017). b. The dry slopes around Enisala fortress, were S. orientalis was collected, are bordered by vast wetland areas, which are partly visible on the left side of the image (photo taken on 26.viii.2016). Photos: L. Székely.
Scopula orientalis is considered to be usually univoltine (mid-May to end July, depending on altitude) (e.g.,
The Romanian specimen of S. orientalis is also among the smallest reported for this species: its 22 mm wing span has been reported only for Turkish specimens, while the usual size is apparently between 24–29 mm (
The mitochondrial DNA (mtDNA) data available have shown that the species of the ornata species-group can be reliably identified based on DNA barcodes, and confirmed the morphology-based identification of S. orientalis from Romania (Fig.
Although S. orientalis is not protected in Europe, the site where this species was found in Romania (Enisala fortress) lies within the territory of the Danube Delta Biosphere Reserve. The rocky steppe habitat lies at the base of Enisala fortress and should be relatively safe from human intervention. This area harbours a series of Lepidoptera species of high zoogeographical value and with extremely limited distributions in Romania and/or Europe (
In conclusion, Scopula orientalis is currently known in Romania only from Enisala fortress, where only one specimen has been found. This situation may nevertheless change if directed research is done (i.e. through dedicated field work, but also re-examination of ornata species-group material stored in collections). Such research could also reveal the presence of the species in other regions of southern Romania, including of course other parts of Dobrogea. For example, if the species is found in southern Dobrogea, this would suggest the presence of S. orientalis in north-eastern Bulgaria as well. The record from Enisala fortress diminishes the geographic gap between the populations from the Balkans and those from southern Ukraine, and also suggests that S. orientalis may be present in the Republic of Moldavia (Fig.
We are grateful to I. Kostjuk, Y. Khalaim, V. Savchuk, S. Beshkov and G. Petranyi for the data and literature provided. We thank subject editor Sven Erlacher, as well as S. Beshkov, P. Huemer, J. Gelbrecht, B. Müller, L. Rákosy and two anonymous reviewers for their comments which improved this study. We thank R. Vila and members of the Butterfly Diversity and Evolution Lab (Barcelona, Spain) for support with DNA sequencing.
Table S1. Specimens and DNA sequences used in this study